LOCUS       sequence-287624-        5218 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Yeast Expression plasmid for SARS-CoV-2 nsp10.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5218)
  TITLE     SARS-CoV-2 protein Yeast expression constructs, tagged, for protein 
            expression and/or purification
REFERENCE   2  (bases 1 to 5218)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"
COMMENT     Sequence Label: pGBW-m4046436
FEATURES             Location/Qualifiers
     source          1..5218
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        10..417
                     /gene="S. cerevisiae TEF1"
                     /label=TEF1 promoter
                     /note="promoter for EF-1-alpha"
     CDS             845..862
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             863..928
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     primer_bind     complement(973..991)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     CDS             1342..2145
                     /codon_start=1
                     /gene="S. cerevisiae URA3"
                     /product="orotidine-5'-phosphate decarboxylase, required 
                     for uracil biosynthesis"
                     /label=URA3
                     /note="yeast auxotrophic marker, counterselectable with 
                     5-fluoroorotic acid (5-FOA)"
                     /translation="MSKATYKERAATHPSPVAAKLFNIMHEKQTNLCASLDVRTTKELL
                     ELVEALGPKICLLKTHVDILTDFSMEGTVKPLKALSAKYNFLLFEDRKFADIGNTVKLQ
                     YSAGVYRIAEWADITNAHGVVGPGIVSGLKQAAEEVTKEPRGLLMLAELSCKGSLSTGE
                     YTKGTVDIAKSDKDFVIGFIAQRDMGGRDEGYDWLIMTPGVGLDDKGDALGQQYRTVDD
                     VVSTGSDIIIVGRGLFAKGRDAKVEGERYRKAGWEAYLRRCGQQN"
     rep_origin      2228..3210
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     terminator      3288..3317
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     rep_origin      3339..3926
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     terminator      complement(3938..3965)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(3989..4180)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(join(4181..4972,4973..5041))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESLRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLATIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGSLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     4804..4823
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5042..5144)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     terminator      complement(5145..5176)
                     /label=tonB terminator
                     /note="bidirectional E. coli tonB-P14 transcription 
                     terminator"
ORIGIN
        1 gcggccgccc cacacaccat agcttcaaaa tgtttctact ccttttttac tcttccagat
       61 tttctcggac tccgcgcatc gccgtaccac ttcaaaacac ccaagcacag catactaaat
      121 ttcccctctt tcttcctcta gggtgtcgtt aattacccgt actaaaggtt tggaaaagaa
      181 aaaagagacc gcctcgtttc tttttcttcg tcgaaaaagg caataaaaat ttttatcacg
      241 tttctttttc ttgaaaattt ttttttttga tttttttctc tttcgatgac ctcccattga
      301 tatttaagtt aataaacggt cttcaatttc tcaagtttca gtttcatttt tcttgttcta
      361 ttacaacttt ttttacttct tgctcattag aaagaaagca tagcaatcta atctaagttt
      421 taattacaaa atgggtaatg caacagaagt gcctgccaat tcaactgtat tatctttctg
      481 tgcttttgct gtagatgctg ctaaagctta caaagattat ctagctagtg ggggacaacc
      541 aatcactaat tgtgttaaga tgttgtgtac acacactggt actggtcagg caataacagt
      601 tacaccggaa gccaatatgg atcaagaatc ctttggtggt gcatcgtgtt gtctgtactg
      661 ccgttgccac atagatcatc caaatcctaa aggattttgt gacttaaaag gtaagtatgt
      721 acaaatacct acaacttgtg ctaatgaccc tgtgggtttt acacttaaaa acacagtctg
      781 taccgtctgc ggtatgtgga aaggttatgg ctgtagttgt gatcaactcc gcgaacccat
      841 gcttctcgtg ccgagaggtt ctgattataa agatcatgat ggtgattata aagatcatga
      901 tattgattat aaagatgatg atgataaata aacaggcccc ttttcctttg tcgatatcat
      961 gtaattagtt atgtcacgct tacattcacg ccctcctccc acatccgctc taaccgaaaa
     1021 ggaaggagtt agacaacctg aagtctaggt ccctatttat tttttttaat agttatgtta
     1081 gtattaagaa cgttatttat atttcaaatt tttctttttt ttctgtacaa agcggccgcc
     1141 gaaccactag tgcactgcag tacacggttt ccttgaaatt tttttgattc ggtaatctcc
     1201 gaacagaagg aagaacgaag gaaggagcac agacttagat tggtatatat acgcatatgt
     1261 agtgttgaag aaacatgaaa ttgcccagta ttcttaaccc aactgcacag aacaaaaacg
     1321 tgcaggaaac gaagataaat catgtcgaaa gctacatata aggaacgtgc tgctactcat
     1381 cctagtcctg ttgctgccaa gctatttaat atcatgcacg aaaagcaaac aaacttgtgt
     1441 gcttcattgg atgttcgtac caccaaggaa ttactggagt tagttgaagc attaggtccc
     1501 aaaatttgtt tactaaaaac acatgtggat atcttgactg atttttccat ggagggcaca
     1561 gttaagccgc taaaggcatt atccgccaag tacaattttt tactcttcga ggacagaaaa
     1621 tttgctgaca ttggtaatac agtcaaattg cagtactctg cgggtgtata cagaatagca
     1681 gaatgggcag acattacgaa tgcacacggt gtggtgggcc caggtattgt tagcggtttg
     1741 aagcaggcgg cagaagaagt aacaaaggaa cctagaggcc ttttgatgtt agcagaattg
     1801 tcatgcaagg gctccctatc tactggagaa tatactaagg gtactgttga cattgcgaag
     1861 agcgacaaag attttgttat cggctttatt gctcaaagag acatgggtgg aagagatgaa
     1921 ggttacgatt ggttgattat gacacccggt gtgggtttag atgacaaggg agatgcattg
     1981 ggtcaacagt atagaaccgt ggatgatgtg gtttctacag gatctgacat tattattgtt
     2041 ggaagaggac tatttgcaaa gggaagggat gctaaggtag agggtgaacg ttacagaaaa
     2101 gcaggctggg aagcatattt gagaagatgc ggccagcaaa actaaaaaac tgtattataa
     2161 gtaaatgcat gtatactaaa ctcacaaatt agagcttcaa tttaattata tcagttatta
     2221 cccgagtaac gaagcatctg tgcttcattt tgtagaacaa aaatgcaacg cgagagcgct
     2281 aatttttcaa acaaagaatc tgagctgcat ttttacagaa cagaaatgca acgcgaaagc
     2341 gctattttac caacgaagaa tctgtgcttc atttttgtaa aacaaaaatg caacgcgaga
     2401 gcgctaattt ttcaaacaaa gaatctgagc tgcattttta cagaacagaa atgcaacgcg
     2461 agagcgctat tttaccaaca aagaatctat acttcttttt tgttctacaa aaatgcatcc
     2521 cgagagcgct atttttctaa caaagcatct tagattactt tttttctcct ttgtgcgctc
     2581 tataatgcag tctcttgata actttttgca ctgtaggtcc gttaaggtta gaagaaggct
     2641 actttggtgt ctattttctc ttccataaaa aaagcctgac tccacttccc gcgtttactg
     2701 attactagcg aagctgcggg tgcatttttt caagataaag gcatccccga ttatattcta
     2761 taccgatgtg gattgcgcat actttgtgaa cagaaagtga tagcgttgat gattcttcat
     2821 tggtcagaaa attatgaacg gtttcttcta ttttgtctct atatactacg tataggaaat
     2881 gtttacattt tcgtattgtt ttcgattcac tctatgaata gttcttacta caattttttt
     2941 gtctaaagag taatactaga gataaacata aaaaatgtag aggtcgagtt tagatgcaag
     3001 ttcaaggagc gaaaggtgga tgggtaggtt atatagggat atagcacaga gatatatagc
     3061 aaagagatac ttttgagcaa tgtttgtgga agcggtattc gcaatatttt agtagctcgt
     3121 tacagtccgg tgcgtttttg gttttttgaa agtgcgtcat cagagcgctt ttggttttca
     3181 aaagcgctct gaagttccta tactttctag ctagagaata ggaacttccc gagcggccgc
     3241 gtgttacaag cttgactcca gcgtaactgg actgcaatca actcactggc tcaccttcac
     3301 gggtgggcct ttcttcggta gaaaatcaaa ggatcttctt gagatccttt ttttctgcgc
     3361 gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat
     3421 caagagctac caactctttt tccgaggtaa ctggcttcag cagagcgcag ataccaaata
     3481 ctgttcttct agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta
     3541 catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc
     3601 ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg
     3661 ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac
     3721 agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg
     3781 taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggta aacgcctggt
     3841 atctttatag tcctgtcggg tttcgccacc tctgacttga gcatcgattt ttgtgatgct
     3901 cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc agaaaggccc acccgaaggt
     3961 gagccaggtg attacatttg ggccctcatc agaggttttc accgtcatca ccgaaacgcg
     4021 cgaggcagct gcggtaaagc tcatcagcgt ggtcgtgaag cgattcacag atgtctgcct
     4081 gttcatccgc gtccagctcg ttgagtttct ccagaagcgt taatgtctgg cttctgataa
     4141 agcgggccat gttaagggcg gttttttcct gtttggtcac ttaccaatgc ttaatcagtg
     4201 aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg
     4261 tgtagataac tacgatgcgg gagggcttac catctggccc cagtgctgca atgataccgc
     4321 gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc gggagggccg
     4381 agcgcagaag tgatcctgca actttatccg cctccatcca gtctattaat tgttgccggg
     4441 aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag
     4501 gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat
     4561 caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc
     4621 cgatcgttgc cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc
     4681 ataattctct tactgtcatg ccatccgtga gatgcttttc tgtgactggt gagtactcaa
     4741 ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac
     4801 gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt
     4861 cggggcgtaa actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc
     4921 gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa
     4981 caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca
     5041 ttttagcttc cttagctcct gaaaatctcg ataactcaaa aaatacgccc ggtagtgatc
     5101 ttatttcatt atggtgaaag ttggaacctc ttacgtgccg atcaagtcaa aagcctccgg
     5161 tcggaggctt ttgactttct gctatggagg tcaggtatga tttaaatggt cagttaga
//
